her2 skbr3 cells  (InvivoGen)

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Expression and secretion levels of miR-19a-3p in breast cancer cells. a) Breast cancer cells express different levels of miR-19a-3p depending on the subtypes, with TNBC (basal) expressing the highest levels compared to luminal B, PAM50 Normal, HER2+, luminal A, and normal cells (TCGA data). The data are shown as box and whisker plots, where the box encompasses the 25th percentile, median, and 75th percentile, and the whiskers extend to 1.5 times the interquartile range; b) the two HER2+ breast cancer cell lines KPL-4 and SKBR3 express higher level of miR-19a-3p than the luminal A MCF-7, consistent with TCGA data; c) the three breast cancer cell lines secrete varying amounts of miR-19a-3p, following an expression pattern similar to that in their parental cells. The results are shown as the mean ± SD of three technical replicates from 3 experimental replicates

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Expressing, Whisker Assay

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Central memory T (T CM ) cell phenotype of CD4 + Th1 cells and immune cell populations in the peripheral blood of patients with metastatic HER2 + breast cancer. a-b) CD4 + Th1 cells expressing high levels of miR-19a-3p developed a central memory T (T CM ) cell phenotype (CD45RO + CCR7 + CD62L +) by the end of the cell culture; c-e) in a small cohort of patients with metastatic HER2 + breast cancer, those with a good prognosis and high serum levels of miR-19a-3p showed a trend of higher percentage of CD4 + Th1 than CD4 + Th2 cells (ratio: IFN-γ + /IL-4 +), although not statistically significant (p = 0.08130) and activated CD4 + T and NK cells circulating in their blood. The results of panel a are shown as the mean ± SD of 3 technical replicates. The results in panel b are presented as the most representative of 3 technical replicates and two experimental replicates. The results in panels c-e are shown as the median with a 95% confidence interval (CI)(Available data from n = 15)

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Expressing, Cell Culture

Journal: Breast Cancer Research : BCR

Article Title: High levels of circulating miR-19a-3p in patients with metastatic HER2 + breast cancer are associated with a favorable prognosis and anti-tumor immune responses

doi: 10.1186/s13058-025-02174-8

Figure Lengend Snippet: Proposed model of TME of HER2 + breast cancer modulated by NK cell-mediated ADCC. The proposed model is based on findings from our current and previous work. Further studies are needed to validate and confirm our hypothetical model. Trastuzumab binds to HER2 + cells and engages CD16 on NK cells, activating them and inducing ADCC. This leads to perforin/granzyme B-mediated apoptosis and death of HER2 + breast cancer cells, as well as the release of miR-19a-3p into the TME and eventually into the bloodstream. ADCCalso stimulates the production and secretion of IFN-γ and T cell-recruiting chemokines from NK cells, which activate DCs, promote their migration to draining lymph nodes, and induce migration of lymphocytes into the TME. This process enables TAA cross-presentation to T cells (both CD4 + and CD8 +) and triggers the production and secretion of IL-12 by DCs. IL-12 and IFN-γ promote the differentiation of CD4 + and CD8 + T cells into CD4 + Th1 cells and CD8 + CTLs and enhance the cytotoxic activity of CD8 + CTLs and NK cells. IL-2 promotes the proliferation of CD4 + Th1 cells, CD8 + CTLs, and NK cells, while IFN-γ from CD4 + Th1 and NK cells further activates DCs (enhancing TAA cross-presentation and cytokine release). The activation and differentiation of CD4 + Th1 cells increase the expression and secretion of miR-19a-3p into the TME and bloodstream. Activated CD8 + CTLs infiltrate tumor tissue and work with NK cells to kill tumor cells, further increasing the release of miR-19a-3p. Additionally, CD4 + Th1 cells differentiate into CD4 + Th1 T CM cells (CD45RO + CCR7 + CD62L +). TAAs from the TME and presented by DCs can maintain the expression and secretion of miR-19a-3p by stimulating CD4 + Th1 T CM cells in draining lymph nodes and contributing to higher serum levels of miR-19a-3p. (This figure was created in BioRender )

Article Snippet: MCF-7 (ER +) and SKBR3 (HER2 +) cells were obtained from the American Type Culture Collection (Manassas, VA, USA), and KPL-4 (HER2 +) cells were provided courtesy of Dr. Junichi Kurebayashi (Kawasaki Medical School, Kurashiki, Japan) [ ].

Techniques: Migration, Activity Assay, Activation Assay, Expressing

Journal: Journal of Medicinal Chemistry

Article Title: Dissecting the Efficacy and Immunogenicity of TLR7 Agonist–Antibody Conjugates through the Lens of Fc Effector Function, Conjugation Strategies, and Linker Cleavability

doi: 10.1021/acs.jmedchem.5c01908

Figure Lengend Snippet: “Industry standard” ISAC design ( Anti-HER2_(Cys)_mcE104 , left graphs) was compared directly with the redesigned cleavable/deglycosylated ISAC ( anti-HER2_(DG/Cys)_mcValCitPABC-E104 , right). (A) HER2+ breast cancer cells (SKBR3) were cocultured for 48 h with mouse macrophage reporter cells, looking for activation of the NFκB pathway; (B) HER2+ breast cancer cells (HCC1954) were cocultured with human PBMCs for 24 h, looking for the release of IFNα; and (C) HER2+ breast cancer cells (HCC1954) were implanted in SCID beige mice beige mice ( n = 5/group), looking for a reduction in tumor volume (IP administration, q5dx3). In each case the HER2 targeted ADC (in green) was compared head-to-head with an isotype (anti-CD20/nontargeted) control ADC (in gray). Note that the right-hand panels of Figures 2A–C represent data previously published by our team, included here for reference.

Article Snippet: Once again, our initial studies involved a coculture of HER2+ SKBR3 cells with the murine macrophage RAW264.7 engineered with both an NFκB and IRF reporter gene (RAW-Dual, Invivogen).

Techniques: Activation Assay, Control

Journal: Journal of Medicinal Chemistry

Article Title: Dissecting the Efficacy and Immunogenicity of TLR7 Agonist–Antibody Conjugates through the Lens of Fc Effector Function, Conjugation Strategies, and Linker Cleavability

doi: 10.1021/acs.jmedchem.5c01908

Figure Lengend Snippet: Assessment of ADCC and ADCP activity for ISACs and naked mAb. Glycosylated analytes are shown with a solid line and deglycosylated analytes are shown with a dotted line. (A) ISACs or mAbs were added to a coculture of Jurkat-Lucia NFAT-CD16a and SKBR3 cells (∼2:1) for 6 h. NFAT activation was assessed by induction of luciferase expression; (B) ISACs or mAbs were treated with a coculture of Jurkat-Lucia NFAT-CD32a and SKBR3 cells (∼2:1) for 6 h. NFAT activation was assessed by induction of luciferase expression.

Article Snippet: Once again, our initial studies involved a coculture of HER2+ SKBR3 cells with the murine macrophage RAW264.7 engineered with both an NFκB and IRF reporter gene (RAW-Dual, Invivogen).

Techniques: Activity Assay, Activation Assay, Luciferase, Expressing

Journal: Journal of Medicinal Chemistry

Article Title: Dissecting the Efficacy and Immunogenicity of TLR7 Agonist–Antibody Conjugates through the Lens of Fc Effector Function, Conjugation Strategies, and Linker Cleavability

doi: 10.1021/acs.jmedchem.5c01908

Figure Lengend Snippet: HER2-targeting immune stimulating ADCs were evaluated in a HCC1954 xenograft model using SCID beige mice. ( n = 7 per group) Both noncleavable (green) and cleavable (blue) designs were evaluated. Dosing was performed at 3mpk by IP administration (q4dx3). (A) Tumor volume (left) and survival (right) of noncleavable ISACs in comparison to the vehicle control; (B) tumor volume (left) and survival (right) of cleavable ISACs in comparison to the vehicle control. Raw growth curves are shown in Supporting Figure S6 . Statistical significance vs the vehicle control was established by Mantel-Cox test (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Once again, our initial studies involved a coculture of HER2+ SKBR3 cells with the murine macrophage RAW264.7 engineered with both an NFκB and IRF reporter gene (RAW-Dual, Invivogen).

Techniques: Comparison, Control

Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Article Title: A coiled coil-based pre-targeting drug delivery system for precise treatment of breast cancer.

doi: 10.1016/j.ejpb.2025.114794

Figure Lengend Snippet: Fig. 4. Visualization of the expression levels and binding patterns of EGFR, Her2 and TROP2 in breast cancer tissues, as well as the specificity of four ADCs targeting these proteins. (a) Microphotographs of mIF of EGFR, Her2 and TROP2 in breast cancer tissues along with corresponding 100x sample regions for each marker. (b) Heatmap shows the expression pattern of EGFR, Her2 and TROP2 in breast cancer tissues. (c) The boxplot represents the average percentages of each marker. (d) Microphotographs of mIF of binding specificities of four ADCs in breast cancer tissues along with corresponding 100x sample regions for each ADC. (e) The per centage of each ADC is illustrated in heatmap. (f) The average percentage of each ADC binding is shown in boxplot.

Article Snippet: TNBC cell lines MDA-MB-468 (ACC 738), MDA-MB231 (ACC 732), MDA-MB-453 (ACC 65) and Hs578T (ACC 781), ER+ breast cancer cell line MCF7 (ACC 115), and Her2+ breast cancer cell lines SKBR3 (ACC 736) and BT474 (ACC 64) were purchased from the Leibniz Institute DSMZ.

Techniques: Expressing, Binding Assay, Marker

Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Article Title: A coiled coil-based pre-targeting drug delivery system for precise treatment of breast cancer.

doi: 10.1016/j.ejpb.2025.114794

Figure Lengend Snippet: Fig. 5. Specific cytotoxicity of pre-targeting complex. (a) Breast cancer cell lines were treated with increasing concentration (10, 20, 40, 80, 160, 320, 640 nM) of scFv-Zip2-Zip1-MMAE, scFv-Erbitux-MMAE, scFv-Sacit-MMAE, combination-Zip1-MMAE, scFv-Zip2-Zip1-SNAP, scFv-SNAP, combination-Zip1-SNAP, Zip1-MMAE or MMAE at 37 ◦C for 72 h, cell lines except SKBR3 were treated with increasing concentration (1.25, 2.5, 5, 10, 20, 40, 80, 160, 320, 640 nM) of scFv-Herceptin- MMAE. (b) SKBR3 was incubated with increasing concentration (0.03125, 0.0625, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16 nM) of scFv-Herceptin-MMAE at 37 ◦C for 72 h. Cell viability was determined by XTT) Kit II, and the experiment was carried out in triplicates for three times. One independent experiment was shown with the mean ± SD.

Article Snippet: TNBC cell lines MDA-MB-468 (ACC 738), MDA-MB231 (ACC 732), MDA-MB-453 (ACC 65) and Hs578T (ACC 781), ER+ breast cancer cell line MCF7 (ACC 115), and Her2+ breast cancer cell lines SKBR3 (ACC 736) and BT474 (ACC 64) were purchased from the Leibniz Institute DSMZ.

Techniques: Concentration Assay, Incubation

Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Article Title: A coiled coil-based pre-targeting drug delivery system for precise treatment of breast cancer.

doi: 10.1016/j.ejpb.2025.114794

Figure Lengend Snippet: Fig. 6. Induction of apoptosis by Trop2-targeting ADCs. (a) Scatter plot of one measurement as an example. (b) Statistic analysis of apoptotic cells including early (Q3) and late (Q2) apoptotic cells inducted by Trop2-, EGFR- and Her2 targeting ADCs. The experiment was carried out independently in triplicates for three times. Data were shown as mean ± SEM. The statistical analysis was carried out using one-way ANOVA and corrected by Tukey’s test (*p＜0.05, **** p＜0.0001).

Article Snippet: TNBC cell lines MDA-MB-468 (ACC 738), MDA-MB231 (ACC 732), MDA-MB-453 (ACC 65) and Hs578T (ACC 781), ER+ breast cancer cell line MCF7 (ACC 115), and Her2+ breast cancer cell lines SKBR3 (ACC 736) and BT474 (ACC 64) were purchased from the Leibniz Institute DSMZ.

Techniques:

Journal: bioRxiv

Article Title: SRSF3 is oncogenic in breast but tumor-suppressive in liver by differential regulation of gene expression

doi: 10.1101/2025.03.14.643315

Figure Lengend Snippet: Srsf3 knockout promotes Htatip2 expression in liver cancer but reduces Htatip2 expression in breast cancer visualized by IGV (A). Srsf3 KD in Hepa 1-6 cells, a murine hepatoma cell line, led to increased expression of Htatip2. Hepa1-6 cells were transfected with 40 nM of Srsf3-specific siRNA (si-Srsf3) or non-targeting siRNA (si-NC) and harvested at 48h upon transfection for analysis. Srsf3 KD efficiency was confirmed by Western blot, with Gapdh serving as a sample loading control (B). (C) Srsf3 KD in Hepa 1-6 cells led to increased expression of Htatip2 examined by TaqMan RT-qPCR in three independent experiments. *, p< 0.05 by Student’s t-test.. (D-E) Knockdown (KD) of Srsf3/SRSF3 promotes exon 11 inclusion in Eif4a2 RNA splicing leading to reduction of eIF4A2 protein in mouse and human breast cancer cells. Total cell RNA was extracted from the cultured individual cell lines transfected with 40 nM of the indicated siRNAs and harvested at 48 h after transfection and used for RT-PCR assays with a forward primer from exon 10 and backward primer from exon 12 (D). Total cell extracts from corresponding cell lines prepared at the 48 h after siRNA transfection were immunoblotted for the protein expression of eIF4A2, SRSF3, and GAPDH with the corresponding antibodies (E).

Article Snippet: Mouse breast cancer cell NF639 expressing c-neu oncogene, human breast cancer cell SKBR3 (HER2+) and MCF7 (ER+) was purchased from ATCC.

Techniques: Knock-Out, Expressing, Transfection, Western Blot, Control, Quantitative RT-PCR, Knockdown, Cell Culture, Reverse Transcription Polymerase Chain Reaction